Genome Sequencing Using PacBio SMRT

Before initiating the genome sequencing process, the quantity and purity of the genomic DNA were determined using an Agilent 2100 Bioanalyzer (Agilent Technologies, Santa Clara, CA, USA). This step was performed to ensure the quality of the genomic DNA for obtaining a complete genome sequence. Genome sequencing was conducted using the PacBio RS II single-molecule real-time (SMRT) sequencing technology from Pacific Biosciences (Menlo Park, CA, USA). SMRTbell library inserts of 20 kb were prepared and sequenced using SMRT cells. Raw sequencing data were generated and subjected to de novo assembly utilizing the hierarchical genome assembly process (HGAP) protocol [19 (link)] and RS HGAP4 Assembly in SMRT analysis software (ver. 2.3; Pacific Biosciences, SMRT Link 4.0.0).